A Reaction-Based Ratiometric Bioluminescent Platform for Point-of-Care and Quantitative Detection Using a Smartphone.
A Reaction-Based Ratiometric Bioluminescent Platform for Point-of-Care and Quantitative Detection Using a Smartphone.
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DOI：
10.1021/acs.analchem.2c05422
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发表时间：
2023-04
影响因子：
7.4
通讯作者：
Junbin Li;Na-Na Wang-Na;Mengyi Xiong;Min Dai;Cheng-Ke Xie;Qianqian Wang;Ke Quan;Yibo Zhou
中科院分区：
文献类型：
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作者：
Junbin Li;Na-Na Wang-Na;Mengyi Xiong;Min Dai;Cheng-Ke Xie;Qianqian Wang;Ke Quan;Yibo Zhou
荧光探针凭借其结构稳定性已成为检测不同分析物的有力工具。然而，对激发源的要求在很大程度上阻碍了它们在即时检测中的适用性，以及在复杂样品中引起自体荧光干扰。本文基于生物发光共振能量转移（BRET）原理，开发了一种基于反应的比率生物发光平台，实现了对分析物的无激发检测。该平台具有模块化设计，由NanoLuc-HaloTag融合物作为能量供体组成，合成荧光探针被生物正交标记为识别部分和能量受体。一旦被靶标激活，荧光探针可以被NanoLuc激发，产生显著的BRET信号，导致发光的明显颜色变化，可以通过智能手机轻松记录和定量分析。作为概念的证明，HOCl的荧光探针被合成以构建生物发光系统。结果表明，该系统显示出恒定的蓝色/红色发射比，其与信号强度无关，允许以高灵敏度（检测限（LOD）= 13 nM）和准确度定量HOCl浓度。鉴于其普遍性，这种基于反应的生物发光平台在即时和定量检测反应性物质方面具有巨大潜力。
Fluorescent probes have emerged as powerful tools for the detection of different analytes by virtue of structural tenability. However, the requirement of an excitation source largely hinders their applicability in point-of-care detection, as well as causing autofluorescence interference in complex samples. Herein, based on bioluminescence resonance energy transfer (BRET), we developed a reaction-based ratiometric bioluminescent platform, which allows the excitation-free detection of analytes. The platform has a modular design consisting of a NanoLuc-HaloTag fusion as an energy donor, to which a synthetic fluorescent probe is bioorthogonally labeled as recognition moiety and energy acceptor. Once activated by the target, the fluorescent probe can be excited by NanoLuc to generate a remarkable BRET signal, resulting in obvious color changes of luminescence, which can be easily recorded and quantitatively analyzed by a smartphone. As a proof of concept, a fluorescent probe for HOCl was synthesized to construct the bioluminescent system. Results demonstrated the system showed a constant blue/red emission ratio which is independent to the signal intensity, allowing the quantification of HOCl concentration with high sensitivity (limit of detection (LOD) = 13 nM) and accuracy. Given the universality, this reaction-based bioluminescent platform holds great potential for point-of-care and quantitative detection of reactive species.